Coding
Part:BBa_K4759082:Design
Designed by: Junyuan Shi Group: iGEM23_Jiangnan-China (2023-10-02)
petH-RBS2-petF(D58F)-linker-GFP1-10
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1249
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1249
Illegal NotI site found at 1022 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1249
Illegal BglII site found at 1560
Illegal BglII site found at 2229
Illegal BamHI site found at 1243 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1249
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1249
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
To rapidly screen the most compatible redox chaperone with Olep, a novel redox chaperone sensor was constructed to measure the interaction force between Olep and the redox chaperone based on bimolecular fluorescence complementation techniques. The sensor uses a superfolded green fluorescent protein (sfGFP), and sfGFP is divided into sfGFP-1-10 and sfGFP-11 without fluorescence. The interaction between sfGFP-1-10 and sfGFP-11 pulls down the distance in space. We performed site-directed mutagenesis of petF (D58F)and ligated GFP1-10 at the C terminus of PetF.
Source
ferredoxin reductase PetH (SEQ ID NO.7) and ferredoxin PetF (SEQ ID NO.8) is from the algae (Synechocystis PCC 6803)